A high-performance liquid immunoaffinity chromatography method for determining transferrin-bound iron in serum.

Area of Science:

• Clinical Chemistry
• Analytical Biochemistry
• Chromatography

Background:

• Current serum iron analysis methods, including the ICSH reference method, are influenced by non-transferrin-bound iron (e.g., ferritin).
• Non-transferrin-bound plasma iron, such as iron citrate in hemochromatosis patients, can be inaccurately measured as serum iron by conventional techniques.
• Existing methods lack specificity, potentially leading to misinterpretation of iron status.

Purpose of the Study:

• To develop and validate a specific method for determining serum transferrin-bound iron (serum t Fe).
• To compare the performance of the novel method against established ICSH and fully automated (FA) methods.
• To enhance the accuracy and reliability of iron status measurements.

Main Methods:

• High-performance liquid immunoaffinity chromatography (HPLAC) was employed to isolate transferrin (Tf) and serum t Fe from interfering substances in serum.
• Interferents were separated using an immunoaffinity column.
• The concentration of isolated t Fe was quantified using a sensitive colorimetric reaction.

Main Results:

• The HPLAC method demonstrated high specificity, with minimal interference from common substances like iron saccharate (5%).
• Excellent precision was achieved, with within-run coefficients of variation (CV) ranging from 0.2-0.4% and between-run CVs from 0.4-1.0%.
• The HPLAC method showed strong correlation with the ICSH method (r=0.9993) and the FA method (r=0.9984).

Conclusions:

• The HPLAC method offers a simple, highly precise, and specific approach for measuring serum transferrin-bound iron (serum t Fe).
• This technique provides a significant advantage over the ICSH and FA methods in terms of specificity.
• The HPLAC method has the potential to improve the accuracy of iron status assessment in clinical practice.